RESUMEN
Bone is one of the most prevalent sites of metastases in various epithelial malignancies, including breast cancer and this metastasis to bone often leads to severe skeletal complications in women due to its osteolytic nature. To address this, we devised a novel drug delivery approach using an Alendronate (ALN) functionalized self-assembled porous crystalsomes for concurrent targeting of Oleanolic acid (OA) and ALN (ALN + OA@NCs) to bone metastasis. Initially, the conjugation of both PEG-OA and OA-PEG-ALN with ALN and OA was achieved, and this conjugation was then self-assembled into porous crystalsomes (ALN + OA@NCs) by nanoemulsion crystallization. The reconstruction of a 3D single particle using transmission electron microscopy ensured the crystalline porous structure of ALN + OA@NCs, was well aligned with characteristic nanoparticle attributes including size distribution, polydispersity, and zeta potential. Further, ALN + OA@NCs showed enhanced efficacy in comparison to OA@NCs suggesting the cytotoxic roles of ALN towards cancer cells, followed by augmentation ROS generation (40.81%), mitochondrial membrane depolarization (57.20%), and induction of apoptosis (40.43%). We found that ALN + OA@NCs facilitated inhibiting osteoclastogenesis and bone resorption followed by inhibited osteolysis. In vivo activity of ALN + OA@NCs in the 4 T1 cell-induced tibia model rendered a reduced bone loss in the treated mice followed by restoring bone morphometric markers which were further corroborated bone-targeting effects of ALN + OA@NCs to reduce RANKL-stimulated osteoclastogenesis. Further, In vivo intravenous pharmacokinetics showed the improved therapeutic profile of the ALN + OA@NCs in comparison to the free drug, prolonging the levels of the drug in the systemic compartment by reducing the clearance culminating the higher accumulation at the tumor site. Our finding proposed that ALN + OA@NCs can effectively target and treat breast cancer metastasis to bone and its associated complications.
Asunto(s)
Alendronato , Neoplasias Óseas , Nanopartículas , Osteólisis , Tibia , Alendronato/administración & dosificación , Alendronato/farmacocinética , Alendronato/química , Animales , Osteólisis/prevención & control , Osteólisis/tratamiento farmacológico , Femenino , Nanopartículas/química , Nanopartículas/administración & dosificación , Tibia/efectos de los fármacos , Tibia/patología , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/secundario , Porosidad , Línea Celular Tumoral , Humanos , Conservadores de la Densidad Ósea/administración & dosificación , Conservadores de la Densidad Ósea/farmacocinética , Ácido Oleanólico/administración & dosificación , Ácido Oleanólico/farmacocinética , Ácido Oleanólico/química , Ácido Oleanólico/análogos & derivados , Ratones Endogámicos BALB C , Ratones , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patologíaRESUMEN
The current study aimed to develop alendronate (ALN)-loaded chitosan nanoparticles (CS-ALN-NPs) for brain delivery via intranasal route. These CS-ALN-NPs reduced the peripheral side effects and released ALN directly to brain. These NPs were formulated through ionic gelation technique by mixing sodium tripolyphosphate (1.5 mg/ml) in ALN-CS (1.75 mg/ml) solution. CS-ALN-NPs attained 135.75 ± 5.80 nm, 0.21 ± 0.013, 23.8 ± 3.69 mV, 72.46 ± 0.879% and 30.92 ± 0.375% mean particle size, PDI, zeta potential, entrapment efficiency and loading capacity, respectively. Furthermore, the TEM and SEM analysis of CS-ALN-NPs, respectively, revealed the particle size in 200 nm range and spherical shape. The in vitro and ex vivo release profile revealed a sustained drug release through CS-ALN-NPs as compared to pure drug solution. Also these NPs acquired a high concentration in mice brain and better pharmacokinetic profile than ALN solution (intranasal) CS-ALN-NPs were then evaluated against intracerebroventricular-streptozotocin (ICV-STZ) induced Alzheimer's disease (AD)-like pathologies in mice. The intranasal CS-ALN-NP altered the ICV-STZ induced neurobehavioral, neurochemical and histopathological changes in mice. These effects were significant to those of ALN solution (intranasal). The neuroprotective potential of CS-ALN-NPs observed in ICV-STZ mice model of AD may be a promising brain-targeted delivery system for AD treatment along with further extensive exploration at both pre-clinical and clinical edge. HIGHLIGHTS CS-ALN-NPs were developed and optimised to overcome the poor pharmacokinetic profile and associated side effects of ALN CS-ALN-NPs showed particle size within 200 nm range as well as controlled and sustained release in in vitro release study These optimised NPs of ALN attained higher brain:blood ratio and better pharmacokinetic profile (Cmax, tmax, AUC) CS-ALN-NPs markedly altered ICV STZ induced impairment in cognitive functions of mice and changes in APP processing, neuroinflammatory cytokines and other biochemical parameters in mice hippocampus.
Asunto(s)
Alendronato/administración & dosificación , Enfermedad de Alzheimer/tratamiento farmacológico , Sistemas de Liberación de Medicamentos , Nanopartículas , Administración Intranasal , Alendronato/farmacocinética , Alendronato/farmacología , Enfermedad de Alzheimer/fisiopatología , Animales , Encéfalo/metabolismo , Encéfalo/fisiopatología , Quitosano/química , Preparaciones de Acción Retardada , Modelos Animales de Enfermedad , Portadores de Fármacos/química , Liberación de Fármacos , Femenino , Masculino , Ratones , Fármacos Neuroprotectores/administración & dosificación , Fármacos Neuroprotectores/farmacocinética , Fármacos Neuroprotectores/farmacología , Tamaño de la Partícula , Estreptozocina , Distribución TisularRESUMEN
The aim of the current research work was to develop sonophoresis-assisted transdermal patches for the treatment of osteoporosis. In the present investigation, we formulated alendronate-chitosan nanoparticles by ionotropic external gelation method. The prepared nanoparticles were found to be smooth and free-flowing. The optimized formulation showed 82.7% of drug release over a period of 12 hours with 99.54% EE, the particle size of 250 nm, PDI 0.22 and zeta potential of 28 mV. The solvent casting evaporation method was used for the development of the patches using HPMC as rate-controlling polymer and dibutyl phthalate as the plasticizer. The optimized patch formulation was found acceptable in terms of physical characteristics (appearance, thickness, folding endurance, weight variation, moisture loss and uptake). The drug content was found to be 99.66±0.9 % with 69.44% of drug permeation through the rat skin. The TP3 formulation had drug content of 99.96% which was the highest among all of the formulations and showed relatively controlled skin permeation of 69.44% over the period of 12 hours. Nearly six-time enhancement of bioavailability was observed when alendronate was used in the nanoparticulate form in transdermal patches used with sonophoresis. Over the period of seven days, the plasma calcium concentration in the rat model was decreased from 16 mg/dl to 4 mg/dl (4 times) in rat groups treated with the transdermal patches containing CS-ALN-NP while the concentration dropped only to 12 mg/dl in case of the transdermal patches containing pure Alendronate. These findings (enhanced skin permeation, enhanced bioavailability and suppression of the plasma calcium level) regarding the transdermal delivery system suggest a promising approach for the treatment of osteoporosis.
Asunto(s)
Alendronato/administración & dosificación , Conservadores de la Densidad Ósea/administración & dosificación , Sistemas de Liberación de Medicamentos/métodos , Osteoporosis/tratamiento farmacológico , Parche Transdérmico , Ondas Ultrasónicas , Administración Cutánea , Alendronato/farmacocinética , Animales , Disponibilidad Biológica , Conservadores de la Densidad Ósea/farmacocinética , Calcio/sangre , Quitosano/química , Modelos Animales de Enfermedad , Liberación de Fármacos , Masculino , Nanopartículas/química , Tamaño de la Partícula , Permeabilidad/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Piel/química , Resultado del TratamientoRESUMEN
pH is a critical indicator of bone physiological function and disease status; however, noninvasive and real-time sensing of bone pH in vivo has been a challenge. Here, we synthesized a bone pH sensor by labeling alendronate with the H+-sensitive dye fluorescein isothiocyanate (Aln-FITC). Aln-FITC showed selective affinity for hydroxyapatite (HAp) rather than other calcium materials. An in vivo biodistribution study showed that Aln-FITC can be rapidly and specifically delivered to rat bones after caudal vein injection, and the fluorescence lasted for at least 12 h. The fluorescence intensity of Aln-FITC binding to HAp linearly decreased when the pH changed from 6 to 12. This finding was further confirmed on bone blocks and perfused bone when the pH changed from 6.8 to 7.4, indicating unique pH-responsive characteristics in the bone microenvironment. Aln-FITC was then preliminarily applied to evaluate the changes in bone pH in a nude mouse acidosis model. Our results demonstrated that Aln-FITC might have the potential for minimally invasive and real-time in vivo bone pH sensing in preclinical studies of bone healing, metabolism, and cancer mechanisms.
Asunto(s)
Alendronato , Huesos , Durapatita , Fluoresceína-5-Isotiocianato , Concentración de Iones de Hidrógeno , Alendronato/análisis , Alendronato/química , Alendronato/farmacocinética , Animales , Huesos/química , Huesos/metabolismo , Durapatita/química , Durapatita/metabolismo , Fluoresceína-5-Isotiocianato/análisis , Fluoresceína-5-Isotiocianato/química , Fluoresceína-5-Isotiocianato/farmacocinética , Humanos , Masculino , Ratones Desnudos , Monitoreo Fisiológico , Imagen Óptica , Ratas , Ratas Sprague-Dawley , Distribución TisularRESUMEN
Researches of biomaterials for osteoporotic bone defects focus on the improvement of its anti-osteoporosis ability, due to osteoporosis is a kind of systemic and long-range bone metabolism disorder. Nevertheless, how to steadily deliver anti-osteoporosis drugs in osteoporotic bone defects is rarely studied. Reported evidences have shown that alendronate (Aln) is known to not only restrain osteoclasts from mediating bone resorption but also stimulate osteoblasts to regenerate bone tissue. Here, we developed an engineered implantable scaffold that could sustainably release Aln for osteoporotic bone defects. Briefly, Aln was added into 2% collagen (Col) solution to form a 5 mg/ml mixture. Then the mixture was filled into pre-designed round models (diameter: 5 mm, height: 2 mm) and crosslinked to obtain engineered Col-Aln scaffolds. The release kinetics showed that Aln was released at an average rate of 2.99 µg/d in the initial 8 days and could sustainably release for 1 month. To detect the repair effects of the Col-Aln scaffolds for osteoporotic defects, the Col and Col-Aln scaffolds were implanted into 5 mm cranial defects in ovariectomized rats. After 3 months, the cranial defects implanted with Col-Aln scaffolds achieved more bone regeneration in defect area (11.74 ± 3.82%) than Col scaffold (5.12 ± 1.15%) (p < .05). Moreover, ovariectomized rats in Col-Aln scaffold group possessed more trabecular bone in femur metaphysis than Col scaffold group as analyzed by Micro-CT. This study demonstrated the engineered Col-Aln scaffold has the potential to repair osteoporotic bone defects and resist bone loss in osteoporosis.
Asunto(s)
Alendronato , Colágeno , Osteoblastos , Osteoporosis , Alendronato/química , Alendronato/farmacocinética , Alendronato/farmacología , Animales , Colágeno/química , Colágeno/farmacología , Preparaciones de Acción Retardada/química , Preparaciones de Acción Retardada/farmacocinética , Preparaciones de Acción Retardada/farmacología , Femenino , Osteoblastos/metabolismo , Osteoblastos/patología , Osteoporosis/tratamiento farmacológico , Osteoporosis/metabolismo , Osteoporosis/patología , Ovariectomía , Ratas , Ratas Sprague-DawleyRESUMEN
Vγ9Vδ2 T cell immunotherapy has been shown to be effective in delaying tumour growth in both pre-clinical and clinical studies. It has been pointed out the importance of the ability of cells to accumulate within tumours and the association with therapeutic efficacy in clinical studies of adoptive T cell transfer. We have previously reported that alendronate liposomes (L-ALD) increase the efficacy of this therapy after localised or systemic injection of γδ T cells in mice, inoculated with ovarian, melanoma, pancreatic or experimental lung metastasis tumour models, respectively. This study aimed to examine the organ biodistribution and tumour uptake of human γδ T cells in subcutaneous (SC), intraperitoneal (IP) or experimental metastatic lung tumours, established in NOD-SCID gamma (NSG) mice using the melanoma cell line A375Pß6.luc. pre-injected with L-ALD. Overall, small variations in blood profiles and organ biodistribution of γδ T cells among the different tumour models were observed. Exceptionally, IP-tumour and experimental metastatic lung-tumour bearing mice pre-injected with L-ALD showed a significant decrease in liver accumulation, and highest uptake of γδ T cells in lungs and tumour-bearing lungs, respectively. Lower γδ T cell count was found in the SC and IP tumours.
Asunto(s)
Alendronato , Inmunoterapia Adoptiva/métodos , Linfocitos Intraepiteliales , Liposomas , Alendronato/administración & dosificación , Alendronato/farmacocinética , Animales , Células Cultivadas , Humanos , Linfocitos Intraepiteliales/citología , Linfocitos Intraepiteliales/metabolismo , Liposomas/administración & dosificación , Liposomas/farmacocinética , Masculino , Ratones , Ratones Endogámicos NOD , Ratones SCID , Distribución TisularRESUMEN
This study aimed to develop an efficient step-by-step osteosarcoma (OS)-targeting liposome system functionalized with a redox-cleavable, bone- and cluster of differentiation 44 (CD44)-dual-targeting polymer. Furthermore, the effect of coadministration of a tumor-penetrating peptide, internalizing RGD (iRGD), was investigated. First, a bone-targeting moiety, alendronate (ALN), was conjugated with hyaluronic acid (HA), a ligand for CD44. This ALN-HA conjugate was coupled with DSPE-PEG2000-COOH through a bioreducible disulfide linker (-SS-) to obtain a functionalized lipid, ALN-HA-SS-L, to be postinserted into preformed liposomes loaded with doxorubicin (DOX). The roles of ALN, HA, and the redox sensitivity of the ALN-HA-SS-L liposomes (ALN-HA-SS-L-L) in the anti-OS effect were critically evaluated against various reference liposomal formulations (with only ALN, HA, or redox sensitivity). ALN-HA-SS-L-L displayed a zeta potential of -26.07 ± 0.32 mV and selectively disassembled in the presence of a reducing agent, 10 mM glutathione, which can be found in cancer cells. Compared to various reference liposomes, ALN-HA-SS-L-L/DOX had significantly higher cytotoxicity to human OS MG-63 cells alongside high and rapid cellular uptake. In the orthotopic OS nude mouse models, ALN-HA-SS-L-L/DOX showed remarkable tumor growth suppression and prolonged survival time. This result was further improved by the coadministration of iRGD. The antitumor effects of various liposomes were ranked in the same order as the degree of tumor biodistribution shown by in vivo/ex vivo imaging: ALN-HA-SS-L-L coadministered with iRGD > ALN-HA-SS-L-L > HA-SS-L-L > HA-L-L > PEG-L> free drug. ALN-HA-SS-L-L/DOX also reduced the cardiotoxicity of DOX and lung metastases. Overall, this study demonstrated that ALN-HA-SS-L-L/DOX, equipped with bone- and CD44-dual-targeting abilities and redox sensitivity, could be a promising OS-targeted therapy. The efficacy could also be augmented by coadministration of iRGD.
Asunto(s)
Alendronato , Neoplasias Óseas , Receptores de Hialuranos/metabolismo , Proteínas de Neoplasias/metabolismo , Osteosarcoma , Alendronato/química , Alendronato/farmacocinética , Alendronato/farmacología , Animales , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/metabolismo , Neoplasias Óseas/patología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Femenino , Humanos , Liposomas , Ratones Endogámicos BALB C , Ratones Desnudos , Osteosarcoma/tratamiento farmacológico , Osteosarcoma/metabolismo , Osteosarcoma/patología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The treatment of metastatic tumors is highly desirable in clinics, which has also increased the interest in the design of nanoscale drug delivery systems. Bone metastasis is one of the most common pathways in the metastasis of breast cancer, and it is also an important cause for tumor recurrence and death. The aryl boronate group, as an acid-labile linker, has been introduced into nano-assemblies in recent years. Especially, as a proteasome inhibitor anticancer drug with a boric acid group, bortezomib can facilitate the formation of pH-sensitive aryl boric acid ester linkage with the catecholic group. In this study, bortezomib-loaded micelles with bone targeting properties were constructed for the treatment of breast cancer bone metastasis. The mixed micelles employed alendronate (ALN) as the bone-targeting ligand and encapsulated bortezomib-catechol conjugates as the cargo. In vitro and in vivo studies showed that compared with free drugs or control micelles, these prodrug micelles (ALN-NP) exhibited many favorable properties such as reduced systemic toxicity and improved therapeutic effects. Therefore, ALN-NP is promising as a nanovehicle for bone-targeting delivery of chemotherapeutic drugs. Furthermore, this study offers a novel strategy combining bone targeting and aryl boronate-based pH-responsive drug release for anti-metastasis therapy.
Asunto(s)
Alendronato/química , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/secundario , Bortezomib/química , Catecoles/química , Sistemas de Liberación de Medicamentos , Micelas , Alendronato/farmacocinética , Alendronato/uso terapéutico , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapéutico , Bortezomib/farmacocinética , Bortezomib/uso terapéutico , Neoplasias de la Mama/patología , Línea Celular , Modelos Animales de Enfermedad , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/uso terapéutico , Femenino , Hemólisis/efectos de los fármacos , Concentración de Iones de Hidrógeno , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Profármacos/química , Profármacos/farmacocinética , Profármacos/uso terapéutico , Inhibidores de Proteasoma/química , Inhibidores de Proteasoma/farmacocinética , Inhibidores de Proteasoma/uso terapéutico , Células RAW 264.7RESUMEN
The scientific objective of this study was to understand the influence of PCL coating on alendronate drug release kinetics in vitro. Our hypothesis was PCL coating would minimize burst release of alendronate from plasma sprayed Mg-doped hydroxyapatite (HA) coated commercially pure titanium (CpTi) samples. In the US alone, over 44 million women and men aged 50 and older are affected by osteoporosis which can lead to replacement and/or revision surgeries. Alendronate is a widely-used drug for treating osteoporosis and would be an ideal drug to be loaded and released from these replacement systems. Initial burst release is a common phenomenon for the most drug loaded devices. To modulate the release kinetics, a biodegradable polymer, polycaprolactone (PCL), coating with slow degradable kinetics was employed. Samples with 2 and 4â¯wt% PCL showed about 34% and 26% release of alendronate within the first 24â¯h, respectively, compared to 75% burst release without any PCL coating. With the addition of a PCL coating, a controlled release kinetics of alendronate was achieved from HA coated titanium implants, which can potentially impact millions of patients worldwide having compromised bone due to osteoporosis.
Asunto(s)
Alendronato , Materiales Biocompatibles Revestidos/química , Durapatita/química , Magnesio/química , Poliésteres/química , Titanio/química , Alendronato/química , Alendronato/farmacocinética , Preparaciones de Acción Retardada/química , Preparaciones de Acción Retardada/farmacocinéticaRESUMEN
Bisphosphonates target and bind avidly to the mineral (hydroxyapatite) found in bone. This targeting ability has been exploited to design and prepare bisphosphonate conjugate prodrugs to deliver a wide variety of drug molecules selectively to bones. It is important that conjugates be stable in the blood stream and that conjugate that is not taken up by bone is eliminated rapidly. The prodrugs should release active drug at a rate appropriate so as to provide efficacy. Radiolabelling is the best method to quantify and evaluate pharmacokinetics, tissue distribution, bone uptake and release of the active drug(s). Recent reports have described bisphosphonate conjugates derived from the antiresorptive drug, alendronic acid and anabolic prostanoid drugs that effectively deliver prostaglandins and prostaglandin EP4 receptor agonists to bone and show enhanced anabolic efficacy and tolerability compared to the drugs alone. These conjugate drugs can be dosed infrequently (weekly or bimonthly) whereas the free drugs must be dosed daily.
Asunto(s)
Alendronato/administración & dosificación , Enfermedades Óseas/tratamiento farmacológico , Huesos/efectos de los fármacos , Difosfonatos/administración & dosificación , Portadores de Fármacos , Profármacos/administración & dosificación , Prostaglandinas/administración & dosificación , Alendronato/efectos adversos , Alendronato/química , Alendronato/farmacocinética , Animales , Enfermedades Óseas/diagnóstico , Enfermedades Óseas/metabolismo , Enfermedades Óseas/fisiopatología , Huesos/metabolismo , Huesos/fisiopatología , Preparaciones de Acción Retardada , Difosfonatos/efectos adversos , Difosfonatos/química , Difosfonatos/farmacocinética , Composición de Medicamentos , Durapatita/metabolismo , Humanos , Profármacos/efectos adversos , Profármacos/química , Profármacos/farmacocinética , Prostaglandinas/efectos adversos , Prostaglandinas/química , Prostaglandinas/farmacocinética , Subtipo EP4 de Receptores de Prostaglandina E/agonistas , Subtipo EP4 de Receptores de Prostaglandina E/metabolismoRESUMEN
Oral bioavailability of the anti-osteoporotic drug alendronate (AL) is limited to ≤ 1% due to unfavorable physicochemical properties. To augment absorption across the gastrointestinal mucosa, an ion pair complex between AL and polyethyleneimine (PEI) was formed and incorporated into nanostructured lipid carriers (NLCs) using a modified solvent injection method. When compared to free AL, ion pairing with PEI increased drug encapsulation efficiency in NLCs from 10% to 87%. Drug release from NLCs measured in vitro using fasted state simulated intestinal fluid, pH 6.5 (FaSSIF-V2) was significantly delayed after PEI complexation. Stability of AL/PEI was pH-dependent resulting in 10-fold faster dissociation of AL in FaSSIF-V2 than measured at pH 7.4. Intestinal permeation properties estimated in vitro across Caco-2 cell monolayers revealed a 3-fold greater flux of AL encapsulated as hydrophobic ion complex in NLCs when compared to AL solution (Papp = 8.43 ± 0.14 × 10-6 cm/s and vs. 2.76 ± 0.42 × 10-6 cm/s). Cellular safety of AL/PEI-containing NLCs was demonstrated up to an equivalent AL concentration of 2.5 mM. These results suggest that encapsulation of AL/PEI in NLCs appears a viable drug delivery strategy for augmenting oral bioavailability of this clinically relevant bisphosphonate drug and, simultaneously, increase gastrointestinal safety.
Asunto(s)
Alendronato/administración & dosificación , Conservadores de la Densidad Ósea/administración & dosificación , Diglicéridos/química , Portadores de Fármacos/química , Lípidos/química , Nanoestructuras/química , Polietileneimina/química , Alendronato/farmacocinética , Conservadores de la Densidad Ósea/farmacocinética , Células CACO-2 , Composición de Medicamentos , Estabilidad de Medicamentos , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Absorción Intestinal , Mucosa Intestinal/metabolismoRESUMEN
A simple, highly sensitive, and robust CE method applied to the determination of alendronate (ALN) was developed from matrices for tissue engineering, characterized by being highly complex systems. The novel method was based on the ALN derivatization with o-phthalaldehyde and 2-mercaptoethanol for direct ultraviolet detection at 254 nm. The BGE consisted of 20 mM sodium borate buffer at pH 10, and the electrophoretic parameters were optimized.The method was validated in terms of specificity, linearity, LOD, LOQ, precision, accuracy, and robustness. The LOD and LOQ obtained were 0.8 and 2.7 µg/mL, respectively. In addition, the method offers higher sensitivity and specificity compared to other CE and HPLC methods using UV-detectors, as well as low cost and simplicity that allowed the rapid and simple quantitation of ALN from bone regeneration matrices.
Asunto(s)
Alendronato/análisis , Portadores de Fármacos/química , Electroforesis Capilar/métodos , Espectrofotometría Ultravioleta/métodos , Alendronato/farmacocinética , Materiales Biocompatibles , Límite de Detección , Modelos Lineales , Reproducibilidad de los ResultadosRESUMEN
Myelodysplastic syndromes (MDS) are a diverse group of bone marrow disorders and clonal hematopoietic stem cell disorders characterized by abnormal blood cells, or reduced peripheral blood cell count. Recent clinical studies on combination therapy of decitabine (DAC) and arsenic trioxide (ATO) have demonstrated synergy on MDS treatment, but the treatment can cause significant side effects to patients. In addition, both drugs have to be administered on a daily basis due to their short half-lives. In addressing key issues of reducing toxic side effects and improving pharmacokinetic profiles of the therapeutic agents, we have developed a new formulation by co-packaging DAC and ATO into alendronate-conjugated bone-targeting nanoparticles (BTNPs). Our pharmacokinetic studies revealed that intravenously administered BTNPs increased circulation time up to 3days. Biodistribution analysis showed that the BTNP facilitated DAC and ATO accumulation in the bone, which is 6.7 and 7.9 times more than untargeted NP. Finally, MDS mouse model treated with BTNPs showed better restoration of complete blood count to normal level, and significantly longer median survival as compared to free drugs or untargeted NPs treatment. Our results support bone-targeted co-delivery of DAC and ATO for effective treatment of MDS.
Asunto(s)
Antineoplásicos/administración & dosificación , Arsenicales/administración & dosificación , Azacitidina/análogos & derivados , Síndromes Mielodisplásicos/tratamiento farmacológico , Nanopartículas/administración & dosificación , Óxidos/administración & dosificación , Alendronato/administración & dosificación , Alendronato/química , Alendronato/farmacocinética , Alendronato/uso terapéutico , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Apoptosis/efectos de los fármacos , Trióxido de Arsénico , Arsenicales/química , Arsenicales/farmacocinética , Arsenicales/uso terapéutico , Azacitidina/administración & dosificación , Azacitidina/química , Azacitidina/farmacocinética , Azacitidina/uso terapéutico , Células de la Médula Ósea/efectos de los fármacos , Huesos/metabolismo , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Decitabina , Ratones Transgénicos , Síndromes Mielodisplásicos/metabolismo , Nanopartículas/química , Nanopartículas/uso terapéutico , Óxidos/química , Óxidos/farmacocinética , Óxidos/uso terapéutico , Fosfatidiletanolaminas/administración & dosificación , Fosfatidiletanolaminas/química , Fosfatidiletanolaminas/farmacocinética , Fosfatidiletanolaminas/uso terapéutico , Polietilenglicoles/administración & dosificación , Polietilenglicoles/química , Polietilenglicoles/farmacocinética , Polietilenglicoles/uso terapéutico , Distribución TisularRESUMEN
Bone is the most common organ affected by metastatic breast cancer. Targeting cancers within the bone remains a great challenge due to the inefficient delivery of therapeutic to bone. In this study, a polyethylene glycol (PEG) coated nanoparticles (NPs) made of a Zn2+ coordination polymer was linked with a bone seeking moiety, alendronate (ALN), to deliver cisplatin prodrug (DSP) to the bone. The particle sizes of this novel system, DSP-Zn@PEG-ALN NPs, were regulated by adjusting the volume ratio of water phase to oil phase in microemulsion. It was small enough (about 55nm) to extravasate through the clefts (80nm) of the bone's sinusoidal capillaries and localize into metastatic bones. DSP-Zn@PEG-ALN NPs showed much higher affinity for hydroxyapatite in vitro and bone in vivo than non-targeted DSP-Zn@PEG NPs and cisplatin. In addition, the in vivo biodistribution studies demonstrated that about 4-fold of platinum was delivered to the bone metastatic lesions than that in healthy bones by DSP-Zn@PEG-ALN NPs intravenously. Finally, DSP-Zn@PEG-ALN NPs not only inhibited the tumor growth efficiently but also reduced the osteocalastic bone destruction. Besides, DSP-Zn@PEG-ALN NPs showed significantly reduced toxicity of cisplatin. These results indicate that the DSP-Zn@PEG-ALN NPs have a great potential in enhancing chemotherapeutic efficacy for the treatment of bone metastatic breast cancer.
Asunto(s)
Alendronato/administración & dosificación , Antineoplásicos/administración & dosificación , Cisplatino/administración & dosificación , Portadores de Fármacos/administración & dosificación , Estructuras Metalorgánicas/administración & dosificación , Nanopartículas/administración & dosificación , Zinc/administración & dosificación , Alendronato/química , Alendronato/farmacocinética , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/metabolismo , Neoplasias Óseas/secundario , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cisplatino/química , Cisplatino/farmacocinética , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Liberación de Fármacos , Humanos , Masculino , Estructuras Metalorgánicas/química , Estructuras Metalorgánicas/farmacocinética , Ratones Endogámicos BALB C , Ratones Desnudos , Nanopartículas/química , Polietilenglicoles/administración & dosificación , Polietilenglicoles/química , Polietilenglicoles/farmacocinética , Profármacos/administración & dosificación , Profármacos/química , Profármacos/farmacocinética , Ratas Sprague-Dawley , Ensayos Antitumor por Modelo de Xenoinjerto , Zinc/química , Zinc/farmacocinéticaRESUMEN
The αvß6 integrin receptor has been shown to be overexpressed on many types of cancer cells, resulting in a more pro-invasive and aggressive phenotype, this makes it an attractive target for selective drug delivery. In tumours that over-express the αvß6 receptor, cellular uptake of liposomes can be enhanced using ligand-targeted liposomes. It has previously been shown in both in vitro and in vivo studies that liposomal alendronate (L-ALD) can sensitise cancer cells to destruction by Vγ9Vδ2 T cells. It is hypothesised that by using the αvß6-specific peptide A20FMDV2 as a targeting moiety for L-ALD, the therapeutic efficacy of this therapy can be increased in αvß6 positive tumours. Targeted liposomes (t-L) were formulated and the targeting efficacy of targeted liposomes (t-L) was assessed by cell uptake and cytotoxicity studies in the αvß6 positive cells line A375Pß6. Bio-distribution of both L and t-L were carried out in αvß6 positive (A375Pß6 and PANC0403) and αvß6 negative (A375Ppuro and PANC-1) subcutaneous tumour mouse models. Immuno-compromised mice bearing A375Pß6 experimental metastatic lung tumours were treated with L-ALD or t-L-ALD as monotherapies or in combination with ex vivo-expanded Vγ9Vδ2 T cells. In vitro, αvß6-dependant uptake of t-L was observed, with t-L-ALD being more effective than L-ALD at sensitising A375Pß6 to γδ T cells. Interestingly, t-L-ALD led to slightly higher but not significant reduction in tumour growth compared to L-ALD, when used as monotherapy in vivo. Moreover, both L-ALD and t-L-ALD led to significant reductions in tumour growth when used in combination with γδ T cells in vivo but t-L-ALD offered no added advantage compared to L-ALD.
Asunto(s)
Alendronato/administración & dosificación , Antígenos de Neoplasias/inmunología , Inmunoterapia , Integrinas/inmunología , Neoplasias/terapia , Subgrupos de Linfocitos T/inmunología , Alendronato/farmacocinética , Animales , Línea Celular Tumoral , Supervivencia Celular , Femenino , Humanos , Liposomas , Masculino , Ratones Endogámicos BALB C , Ratones SCID , Neoplasias/inmunología , Neoplasias/metabolismo , Neoplasias/patología , Carga TumoralRESUMEN
In search of an effective and less toxic absorption enhancer, we synthesized primary amine acetylation of generation 2 polyamidoamine (G2 PAMAM) dendrimer (Ac-G2) by the reaction of G2 PAMAM dendrimer with acetic anhydride, and evaluated the effects of Ac-G2 on the intestinal absorption of poorly absorbable water-soluble drugs using an in situ closed-loop method in rats. The results indicated that Ac50-G2 had a greatest absorption enhancing effect for 5(6)-carboxyfluorescein (CF) in various acetylation levels of G2 PAMAM dendrimers. Ac50-G2 with various concentrations (0.1-1.0%, w/v) could significantly improve the intestinal absorption of alendronate, CF, and fluorescein isothiocyanate-labeled dextrans (FD4), although they did not enhance the absorption of macromolecular drug of FD10, and the absorption enhancement effect of Ac50-G2 was concentration-dependent. Furthermore, we examined the intestinal membrane damage with or without Ac50-G2. The results displayed Ac50-G2 at lower concentrations (0.1-0.5%, w/v) did not cause any observed toxic effect to the intestinal membranes. These findings suggested Ac50-G2 at lower concentrations (below 0.5%, w/v) might be promising as an effective and safe absorption enhancers to promote the intestinal absorption of poorly absorbable drugs.
Asunto(s)
Anhídridos Acéticos/química , Dendrímeros/administración & dosificación , Absorción Intestinal/efectos de los fármacos , Acetilación , Alendronato/administración & dosificación , Alendronato/sangre , Alendronato/farmacocinética , Animales , Dendrímeros/química , Dextranos/administración & dosificación , Dextranos/sangre , Dextranos/farmacocinética , Fluoresceína-5-Isotiocianato/administración & dosificación , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/farmacocinética , Fluoresceínas/administración & dosificación , Fluoresceínas/farmacocinética , Colorantes Fluorescentes/administración & dosificación , Colorantes Fluorescentes/farmacocinética , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/metabolismo , Masculino , Ratas Wistar , Solubilidad , Agua/químicaRESUMEN
A chitosan-based microsphere delivery system has been fabricated for controlled release of alendronate (AL). The present study aimed to incorporate the chitosan/hydroxyapatite microspheres-loaded with AL (CH/nHA-AL) into poly(L-lactic acid)/nanohydroxyapatite (PLLA/nHA) matrix to prepare a novel microspheres-scaffold hybrid system (CM-ALs) for drug delivery and bone tissue engineering application. The characteristics of CM-ALs scaffolds containing 10% and 20% CH/nHA-AL were evaluated in vitro, including surface morphology and porosity, mechanical properties, drug release, degradation, and osteogenic differentiation. The in vivo bone repair for large segmental radius defects (1.5 cm) in a rabbit model was evaluated by radiography and histology. In vitro study showed more sustained drug release of CM-AL-containing scaffolds than these of CM/nHA-AL and PLLA/nHA/AL scaffolds, and the mechanical and degradation properties of CM-ALs (10%) scaffolds were comparable to that of PLLA/nHA control. The osteogenic differentiation of adipose-derived stem cells (ASCs) was significantly enhanced as indicated by increased alkaline phosphates (ALP) activity and calcium deposition. In vivo study further showed better performance of CM-ALs (10%) scaffolds with complete repair of large-sized bone defects within 8 weeks. A microspheres-scaffold-based release system containing AL-encapsulated chitosan microspheres was successfully fabricated in this study. Our results suggested the promising application of CM-ALs (10%) scaffolds for drug delivery and bone tissue engineering.
Asunto(s)
Alendronato/administración & dosificación , Regeneración Ósea/efectos de los fármacos , Sustitutos de Huesos/química , Sistemas de Liberación de Medicamentos/métodos , Andamios del Tejido , Alendronato/farmacocinética , Animales , Células Cultivadas , Quitosano/química , Durapatita/química , Técnicas In Vitro , Ácido Láctico/química , Microesferas , Osteogénesis/efectos de los fármacos , Conejos , Ingeniería de Tejidos/métodosRESUMEN
A novel multifunctional Ag2 S quantum dot (QD)-based nanomedicine of alendronate (Ald)/doxorubicin (DOX)@Ag2 S is developed for highly effective bone tumor therapy in an orthotopic model. The bone-targeting and osteolysis inhibition of Ald and the chemotherapeutic effect of DOX on the bone tumor are in situ visualized by Ag2 S QDs with emission in the second near-infrared window.
Asunto(s)
Antineoplásicos/farmacología , Neoplasias Óseas/tratamiento farmacológico , Neoplasias Óseas/secundario , Sistemas de Liberación de Medicamentos , Osteólisis/tratamiento farmacológico , Puntos Cuánticos , Alendronato/farmacocinética , Alendronato/farmacología , Animales , Neoplasias Óseas/diagnóstico por imagen , Neoplasias Óseas/fisiopatología , Huesos/diagnóstico por imagen , Huesos/patología , Huesos/fisiopatología , Bovinos , Línea Celular Tumoral , Relación Dosis-Respuesta a Droga , Doxorrubicina/farmacocinética , Doxorrubicina/farmacología , Evaluación Preclínica de Medicamentos , Humanos , Macrófagos/efectos de los fármacos , Macrófagos/patología , Macrófagos/fisiología , Ratones , Nanomedicina , Trasplante de Neoplasias , Osteoclastos/efectos de los fármacos , Osteoclastos/patología , Osteoclastos/fisiología , Osteólisis/diagnóstico por imagen , Osteólisis/patología , Osteólisis/fisiopatología , Puntos Cuánticos/uso terapéutico , Compuestos de Plata/farmacocinética , Compuestos de Plata/farmacologíaRESUMEN
Severe osteoporotic diseases, such as Paget's disease, Osteogenesis Imperfecta, and Legg Calve Perthes disease, lack treatments that address the pathobiology of the diseases, as well as, long-term and prospective studies. Bisphosphonates, which are known to dramatically hinder the viability of osteoclast cells, along with gold nanoparticles (GNP) are a potential theranostic for osteoporotic diseases. We evaluated GNP functionalized with two different bisphosphonates, namely, alendronate and pamidronate. RANKL differentiated murine pre-osteoclasts (Raw 264.7) and murine osteoblasts (7F2) were treated with varying concentrations ranging from 0.1-5 µM of free and GNP bound bisphosphonates. GNPs with an average size of â¼15 nm were functionalized with alendronate and pamidronate through surface modification by self-assembly. MTT viability assay results show no changes in viability of the osteoclasts when treated with free bisphosphonates in the range of 1-5 µM, but significant decrease on treatment with functionalized GNP at concentrations above the range of 0.1-1 µM depending on the bisphosphonate. Osteoblast cell viability is maintained at all but the highest concentrations used. Qualitative and quantitative characterization by Western Blot for RANKL expression in the osteoblast cell line shows that expression is largely maintained. These results provide a basis for methods that use bisphosphonate functionalized GNP in the treatment of osteoporotic bone diseases. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 21-29, 2017.
Asunto(s)
Alendronato/farmacocinética , Difosfonatos/farmacología , Oro , Nanopartículas del Metal/química , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Animales , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Oro/química , Oro/farmacología , Ratones , Osteoblastos/citología , Osteoclastos/citología , Pamidronato , Células RAW 264.7RESUMEN
Alendronate (ALN) is known as an anti-resorptive drug for the treatment of osteoporosis. Recently, ALN was found to stimulate osteogenic differentiation in mesenchymal stem cells and enhance new bone formation in calvarial bone defects. Previous in vitro and in vivo studies found that the effective concentration of ALN was approximately 1-10 µm. In the present study, a poly (lactic-co-glycolic acid) (PLGA) cross-linked ALN (PLGA-ALN) with a short-term controlled-release property for local application to enhance bone repair was developed. An in vitro drug-release kinetic test showed that PLGA-ALN microspheres released an effective concentration (50-100 nm) of ALN for 9 days. The effect of PLGA-ALN on bone repair was tested in a rat femoral bone defect model. The biomechanical study results showed that the maximal strength, stiffness and energy absorption were significantly increased in the PLGA-ALN group compared with the PLGA group. The microstructure of the newly formed bone at the defect site was analysed using microcomputed tomography. The PLGA-ALN group significantly improved the trabecular bone volume at the defect site compared with the PLGA group. The fibril collagen and immunolocalized bone morphogenetic protein 2 were evident in the newly formed trabecular bone in the PLGA-ALN group. Local use of newly developed PLGA-ALN-enhanced bone repair was attributable to increasing bone matrix formation, which improved the ultrastructure of the newly formed bone and thus increased the biomechanical properties of the repaired bone. It is suggested that PLGA-ALN may be a potential bone graft substitute to enhance bone repair. Copyright © 2016 John Wiley & Sons, Ltd.